RT Journal Article
SR Electronic
T1 LED-based interference-reflection microscopy combined with optical tweezers for quantitative three-dimensional single microtubule imaging
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 277632
DO 10.1101/277632
A1 Steve Simmert
A1 Mohammad Kazem Abdosamadi
A1 Gero Hermsdorf
A1 Erik Schäffer
YR 2018
UL http://biorxiv.org/content/early/2018/03/07/277632.abstract
AB Optical tweezers combined with various microscopy techniques are a versatile tool for single-molecule force spectroscopy. However, some combinations may compromise measurements. Here, we combined optical tweezers with total-internal-reflection-fluorescence (TIRF) and interference-reflection microscopy (IRM). Using a light-emitting diode (LED) for IRM illumination, we show that single microtubules can be resolved with high contrast. Furthermore, we converted the IRM interference pattern of an upward bent microtubule to its three-dimensional (3D) profile calibrated against the optical tweezers and evanescent TIRF field. In general, LED-based IRM is a powerful method for high-resolution 3D microscopy.OCIS codes (180.3170) Interference microscopy; (120.4570) Optical design of instruments (350.4855); Optical tweezers or optical manipulation.